title: An in vitro model of the horse gut microbiome enables identification of lactate-utilizing bacteria that differentially respond to starch induction.
abstract: Laminitis is a chronic, crippling disease triggered by the sudden influx of dietary starch. Starch reaches the hindgut resulting in enrichment of lactic acid bacteria, lactate accumulation, and acidification of the gut contents. Bacterial products enter the bloodstream and precipitate systemic inflammation. Hindgut lactate levels are normally low because specific bacterial groups convert lactate to short chain fatty acids. Why this mechanism fails when lactate levels rapidly rise, and why some hindgut communities can recover is unknown. Fecal samples from three adult horses eating identical diets provided bacterial communities for this in vitro study. Triplicate microcosms of fecal slurries were enriched with lactate and/or starch. Metabolic products (short chain fatty acids, headspace gases, and hydrogen sulfide) were measured and microbial community compositions determined using Illumina 16S rRNA sequencing over 12-hour intervals. We report that patterns of change in short chain fatty acid levels and pH in our in vitro system are similar to those seen in in vivo laminitis induction models. Community differences between microcosms with disparate abilities to clear excess lactate suggest profiles conferring resistance of starch-induction conditions. Where lactate levels recover following starch induction conditions, propionate and acetate levels rise correspondingly and taxa related to Megasphaeraelsdenii reach levels exceeding 70% relative abundance. In lactate and control cultures, taxa related to Veillonellamontpellierensis are enriched as lactate levels fall. Understanding these community differences and factors promoting the growth of specific lactate utilizing taxa may be useful to prevent acidosis under starch-induction conditions.
authors.affiliation: Department of Microbiology, University of Massachusetts, Amherst, Massachusetts, United States of America.
authors.forename: Amy S
authors.initials: AS
authors.lastname: Biddle
authors.forename: Samuel J
authors.initials: SJ
authors.lastname: Black
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Blanchard
mesh_terms: D019342:Acetic Acid / Q000378:metabolism; D000140:Acidosis, Lactic / Q000382:microbiology; D000821:Animal Feed; D000818:Animals; D058491:Bacterial Load; D002432:Cecum / Q000378:metabolism / Q000382:microbiology; D003106:Colon / Q000378:metabolism / Q000382:microbiology; D005243:Feces / Q000382:microbiology; D006736:Horses; D006863:Hydrogen-Ion Concentration; D019344:Lactic Acid / Q000378:metabolism*; D045851:Megasphaera / Q000302:isolation & purification / Q000378:metabolism*; D064307:Microbiota / Q000502:physiology*; D008954:Models, Biological; D011422:Propionates / Q000378:metabolism; D012336:RNA, Ribosomal, 16S / Q000032:analysis* / Q000235:genetics; D013213:Starch / Q000378:metabolism*; D014678:Veillonella / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D011422:Propionates; D012336:RNA, Ribosomal, 16S; D019344:Lactic Acid; D013213:Starch; C029658:propionic acid; D019342:Acetic Acid